Tocotrienol modulates crucial lipid metabolism-related genes in differentiated 3T3-L1 preadipocytes.
Obesity and other lipid metabolism-related diseases have become more prevalent in recent years due to drastic lifestyle changes and dietary patterns. Unsaturated vitamin E, tocotrienol (T3), represents one of the most fascinating naturally occurring compounds that has the potential to influence a broad range of mechanisms underlying abnormal lipid metabolism processes. However, its efficacy and mechanism have been uncertain due to scarcity of data concerning the effect of T3 on lipid metabolism. In this study, we report a series of fascinating experimental findings on how T3 affects lipid metabolism in differentiated 3T3-L1 preadipocytes. Treatment with T3 (25 μM), especially δ and γ isomers, inhibited the accumulation of triglyceride and lipid droplets in differentiated 3T3-L1 cells. This manifestation was supported by mRNA and protein expression of crucial lipid metabolism-related genes. The present study provides a novel set of data pertaining to the possibility of T3 as an anti-metabolic disorder agent.


Global Reach 2018 Heightened α-Adrenergic Signaling Impairs Endothelial Function During Chronic Exposure to Hypobaric Hypoxia.
Chronic exposure to hypoxia is associated with elevated sympathetic nervous activity and reduced vascular function in lowlanders, and Andean highlanders suffering from excessive erythrocytosis (EE); however, the mechanistic link between chronically elevated sympathetic nervous activity and hypoxia-induced vascular dysfunction has not been determined. To determine the impact of heightened sympathetic nervous activity on resistance artery endothelial-dependent dilation (EDD), and endothelial-independent dilation, in lowlanders and Andean highlanders with and without EE. We tested healthy lowlanders (n=9) at sea level (344 m) and following 14 to 21 days at high altitude (4300 m), and permanent Andean highlanders with (n=6) and without (n=9) EE at high altitude. Vascular function was assessed using intraarterial infusions (3 progressive doses) of acetylcholine (ACh; EDD) and sodium nitroprusside (endothelial-independent dilation) before and after local α+β adrenergic receptor blockade (phentolamine and propranolol). Intraarterial blood pressure, heart rate, and simultaneous brachial artery diameter and blood velocity were recorded at rest and during drug infusion. Changes in forearm vascular conductance were calculated. The main findings were (1) chronic hypoxia reduced EDD in lowlanders (changes in forearm vascular conductance from sea level: ACh1: -52.7±19.6%, ACh2: -25.4±38.7%, ACh3: -35.1±34.7%, all P≤0.02); and in Andeans with EE compared with non-EE (changes in forearm vascular conductance at ACh3: -36.4%, P=0.007). Adrenergic blockade fully restored EDD in lowlanders at high altitude, and normalized EDD between EE and non-EE Andeans. (2) Chronic hypoxia had no effect on endothelial-independent dilation in lowlanders, and no differences were detected between EE and non-EE Andeans; however, EID was increased in the non-EE Andeans after adrenergic blockade (P=0.012), but this effect was not observed in the EE Andeans. These data indicate that chronic hypoxia reduces EDD via heightened α-adrenergic signaling in lowlanders and in Andeans with EE. These vascular mechanisms have important implications for understanding the physiological consequences of acute and chronic high altitude adaptation.


Oxidation of Phenolic Aldehydes by Ozone and Hydroxyl Radicals at the Air-Solid Interface.
Biomass burning emissions contain abundant phenolic aldehydes (e.g., syringaldehyde, vanillin, and 4-hydroxybenaldehyde) that are oxidized during atmospheric transport, altering the physicochemical properties of particulates. Herein, the oxidative processing of thin films made of syringaldehyde, vanillin, and 4-hydroxybenaldehyde is studied at the air-solid interface under a variable O3(g) molar ratio (410 ppbv-800 ppmv) and relative humidity (0-90%). Experiments monitored the absorption changes of C=C, C=O, and -COOH vibration changes during the oxidation of thin films by transmission Fourier transform infrared spectroscopy (FTIR). Selected spectroscopic features of aromatic ring cleavage by O3(g) revealed the production of carboxylic acids. Instead, monitoring O-H stretching provided a comparison of a hydroxylation channel from in situ produced hydroxyl radical. The overall oxidation reactivity trend syringaldehyde > vanillin > 4-hydroxybenzladehyde can be explained based on the additional electron density from methoxide substituents to the ring. The reactive uptake coefficient of O3(g) increases for higher relative humidity, e.g., for syringaldehyde by 18 and 215 times at 74% and 90% relative humidity (RH), respectively, as compared to dry conditions. A Langmuir-Hinshelwood mechanism fits well the kinetics of oxidation under a variable O3(g) molar ratio at 74% RH, providing useful information that should be included in atmospheric chemistry models.


Papillomavirus capsid protein expression in Escherichia coli: purification and assembly of HPV11 and HPV16 L1.
The L1 major capsid proteins of human papillomavirus (HPV) types 11 and 16 were purified and analyzed for structural integrity and in vitro self-assembly. Proteins were expressed in Escherichia coli as glutathione-S-transferase-L1 (GST-L1) fusions and purified to near homogeneity as pentamers (equivalent to viral capsomeres), after thrombin cleavage from the GST moiety and removal of tightly associated GroEL protein. Sequences at the amino and carboxy termini contributing to formation of L1 pentamers and to in vitro capsid assembly were identified by deletion analysis. For both HPV11 and HPV16 L1, up to at least ten residues could be deleted from the amino terminus (Delta N10) and 30 residues from the carboxy terminus (Delta C30) without affecting pentamer formation. The HPV16 pentamers assembled into relatively regular, 72-pentamer shells ("virus-like particles" or VLPs) at low pH, with the exception of HPV16 L1 Delta N10, which assembled into a 12-pentamer, T=1 capsid (small VLP) under all conditions tested. The production of large quantities of assembly-competent L1, using the expression and purification protocol described here, has been useful for crystallographic analysis, and will be valuable for studies of virus-receptor interactions and potentially for vaccine design.


Entropy-based grid approach for handling outliers: a case study to environmental monitoring data.
Grid-based approaches render an efficient framework for data clustering in the presence of incomplete, inexplicit, and uncertain data. This paper proposes an entropy-based grid approach (EGO) for outlier detection in clustered data. The given hard clusters obtained from a hard clustering algorithm, EGO uses entropy on the dataset as a whole or on an individual cluster to detect outliers. EGO works in two steps: explicit outlier detection and implicit outlier detection. Explicit outlier detection is concerned with those data points that are isolated in the grid cells. They are either far from the dense region or maybe a nearby isolated data point and therefore declared as an explicit outlier. Implicit outlier detection is associated with the detection of outliers that are perplexedly deviated from the normal pattern. The determination of such outliers is achieved using entropy change of the dataset or a specific cluster for each deviation. The elbow based on the trade-off between entropy and object geometries optimizes the outlier detection process. Experimental results on CHAMELEON datasets and other similar datasets suggested that the proposed approach(es) detect the outliers more precisely and extend the capability of outliers detection to an additional 4.5% to 8.6%. Moreover, the resultant clusters became more precise and compact when the entropy-based gridding approach is applied on top of hard clustering algorithms. The performance of the proposed algorithms is compared with well-known outlier detection algorithms, including DBSCAN, HDBSCAN, RE3WC, LOF, LoOP, ABOD, CBLOF and HBOS. Finally, a case study for detecting outliers in environmental data has been carried out using the proposed approach and results are generated on our synthetically prepared datasets. The performance shows that the proposed approach may be an industrial-oriented solution to outlier detection in environmental monitoring data.


Substantial out-of-pocket expenditure on maternity care practitioner consultations and treatments during pregnancy: estimates from a nationally-representative sample of pregnant women in Australia.
A wide range of health care options are utilised by pregnant women in Australia. The out-of-pocket costs of maternity care in Australia vary depending on many factors including model of care utilised, health insurance coverage, and women's decision to access health services outside of conventional maternity care provision. Women from the 1973-78 cohort of the Australian Longitudinal Study on Women's Health (ALSWH) who identified as pregnant or as recently having given birth in 2009 were invited to complete a sub-study questionnaire investigating health service utilisation during their most recent pregnancy. A total of 1,835 women agreed to participate in the sub-study. The majority of women (99.8%) consulted with a conventional health care practitioner during pregnancy, 49.4% consulted with a complementary and alternative medicine practitioner at least once during pregnancy and 89.6% of the women used a complementary and alternative medicine product. Women reported an average of AUD$781.10 in out-of-pocket expenses for consultations with conventional health care practitioners, AUD$185.40 in out-of-pocket expenses for consultations with complementary and alternative medicine practitioners and AUD$179.60 in out-of-pocket expenses for complementary and alternative medicine products. From the study data we estimate Australian pregnant women spend over AUD$337 M on out-of-pocket health services. While the majority of pregnant women in Australia may obtain health services via the publically-funded health care system and/or private health insurance coverage, our analysis identifies substantial out-of-pocket expenditure for health care by pregnant women - a trend in public spending for maternity care of importance to policy makers, health administrators, and health professionals.


Structure-function features of a Mycoplasma glycolipid synthase derived from structural data integration, molecular simulations, and mutational analysis.
Glycoglycerolipids are structural components of mycoplasma membranes with a fundamental role in membrane properties and stability. Their biosynthesis is mediated by glycosyltransferases (GT) that catalyze the transfer of glycosyl units from a sugar nucleotide donor to diacylglycerol. The essential function of glycolipid synthases in mycoplasma viability, and the absence of glycoglycerolipids in animal host cells make these GT enzymes a target for drug discovery by designing specific inhibitors. However, rational drug design has been hampered by the lack of structural information for any mycoplasma GT. Most of the annotated GTs in pathogenic mycoplasmas belong to family GT2. We had previously shown that MG517 in Mycoplasma genitalium is a GT-A family GT2 membrane-associated glycolipid synthase. We present here a series of structural models of MG517 obtained by homology modeling following a multiple-template approach. The models have been validated by mutational analysis and refined by long scale molecular dynamics simulations. Based on the models, key structure-function relationships have been identified: The N-terminal GT domain has a GT-A topology that includes a non-conserved variable region involved in acceptor substrate binding. Glu193 is proposed as the catalytic base in the GT mechanism, and Asp40, Tyr126, Tyr169, Ile170 and Tyr218 define the substrates binding site. Mutation Y169F increases the enzyme activity and significantly alters the processivity (or sequential transferase activity) of the enzyme. This is the first structural model of a GT-A glycoglycerolipid synthase and provides preliminary insights into structure and function relationships in this family of enzymes.


[Psychometric properties of the Addenbrooke's Cognitive Examination III (ACE-III) for the detection of dementia].
The Addenbrooke's Cognitive Examination III (ACE-III) is widely used for evaluation of dementia. To assess the psychometric properties of ACE-III, analyzing its factor structure, its reliability (from an Item Response Theory [TRI] model) and its diagnostic usefulness. We studied 1101 older people without cognitive impairment and 63 currently having a diagnosis and receiving treatment for dementia. The presence of two factors for the Attention subscale (Orientation and Attention, separately) was suggested. The factorial analysis showed adequate adjustment in all the subscales, except for the new Attention subscale. In the TRI analysis, the Attention subscale presented a greater number of items with lack of fit compared to the other subscales. Using a proposed threshold of 66 points or less to identify cognitive impairment related to dementia, a sensitivity of 0.97 and specificity of 0.81 was obtained. ACE-III is a valid, reliable, and useful measure for the clinical detection of dementia. The combined use of Orientation and Memory subscales is proposed as an alternative and time-saving ACE-III indicator.


Advanced glycation end products and vascular inflammation: implications for accelerated atherosclerosis in diabetes.
The formation of advanced glycation end products (AGEs) is an important biochemical abnormality that accompanies diabetes mellitus and, likely, inflammation in general. Here we summarize and discuss recent studies indicating that the effects of AGEs on vessel wall homeostasis may account for the rapidly progressive atherosclerosis associated with diabetes mellitus. Driven by hyperglycemia and oxidant stress, AGEs form to a greatly accelerated degree in diabetes. Within the vessel wall, collagen-linked AGEs may "trap" plasma proteins, quench nitric oxide (NO) activity and interact with specific receptors to modulate a large number of cellular properties. On plasma low density lipoproteins (LDL), AGEs initiate oxidative reactions that promote the formation of oxidized LDL. Interaction of AGEs with endothelial cells as well as with other cells accumulating within the atherosclerotic plaque, such as mononuclear phagocytes and smooth muscle cells (SMCs), provides a mechanism to augment vascular dysfunction. Specifically, the interaction of AGEs with vessel wall components increases vascular permeability, the expression of procoagulant activity and the generation of reactive oxygen species (ROS), resulting in increased endothelial expression of endothelial leukocyte adhesion molecules. AGEs potently modulate initiating steps in atherogenesis involving blood-vessel wall interactions, triggering an inflammatory-proliferative process and, furthermore, critically contribute to propagation of inflammation and vascular perturbation in established disease. Thus, a better understanding of the biochemical mechanisms by which AGEs contribute to such processes in the vessel wall could be relevant to devise preventive and therapeutic strategies for diabetic atherosclerosis.


Association Between Titin Loss-of-Function Variants and Early-Onset Atrial Fibrillation.
Atrial fibrillation (AF) is the most common arrhythmia affecting 1% of the population. Young individuals with AF have a strong genetic association with the disease, but the mechanisms remain incompletely understood. To perform large-scale whole-genome sequencing to identify genetic variants related to AF. The National Heart, Lung, and Blood Institute's Trans-Omics for Precision Medicine Program includes longitudinal and cohort studies that underwent high-depth whole-genome sequencing between 2014 and 2017 in 18 526 individuals from the United States, Mexico, Puerto Rico, Costa Rica, Barbados, and Samoa. This case-control study included 2781 patients with early-onset AF from 9 studies and identified 4959 controls of European ancestry from the remaining participants. Results were replicated in the UK Biobank (346 546 participants) and the MyCode Study (42 782 participants). Loss-of-function (LOF) variants in genes at AF loci and common genetic variation across the whole genome. Early-onset AF (defined as AF onset in persons <66 years of age). Due to multiple testing, the significance threshold for the rare variant analysis was P = 4.55 × 10-3. Among 2781 participants with early-onset AF (the case group), 72.1% were men, and the mean (SD) age of AF onset was 48.7 (10.2) years. Participants underwent whole-genome sequencing at a mean depth of 37.8 fold and mean genome coverage of 99.1%. At least 1 LOF variant in TTN, the gene encoding the sarcomeric protein titin, was present in 2.1% of case participants compared with 1.1% in control participants (odds ratio [OR], 1.76 [95% CI, 1.04-2.97]). The proportion of individuals with early-onset AF who carried a LOF variant in TTN increased with an earlier age of AF onset (P value for trend, 4.92 × 10-4), and 6.5% of individuals with AF onset prior to age 30 carried a TTN LOF variant (OR, 5.94 [95% CI, 2.64-13.35]; P = 1.65 × 10-5). The association between TTN LOF variants and AF was replicated in an independent study of 1582 patients with early-onset AF (cases) and 41 200 control participants (OR, 2.16 [95% CI, 1.19-3.92]; P = .01). In a case-control study, there was a statistically significant association between an LOF variant in the TTN gene and early-onset AF, with the variant present in a small percentage of participants with early-onset AF (the case group). Further research is necessary to understand whether this is a causal relationship.


Towards zero-threshold optical gain using charged semiconductor quantum dots.
Colloidal semiconductor quantum dots are attractive materials for the realization of solution-processable lasers. However, their applications as optical-gain media are complicated by a non-unity degeneracy of band-edge states, because of which multiexcitons are required to achieve the lasing regime. This increases the lasing thresholds and leads to very short optical gain lifetimes limited by nonradiative Auger recombination. Here, we show that these problems can be at least partially resolved by employing not neutral but negatively charged quantum dots. By applying photodoping to specially engineered quantum dots with impeded Auger decay, we demonstrate a considerable reduction of the optical gain threshold due to suppression of ground-state absorption by pre-existing carriers. Moreover, by injecting approximately one electron per dot on average, we achieve a more than twofold reduction in the amplified spontaneous emission threshold, bringing it to the sub-single-exciton level. These measurements indicate the feasibility of 'zero-threshold' gain achievable by completely blocking the band-edge state with two electrons.


Minimally invasive treatment of fecal incontinence and constipation in children.
The capacity for voluntary bowel movements and thus fecal incontinence may be limited in children born with anorectal anomalies and Hirschsprung's disease, as well as for many patients with spinal problems. Some of these patients have severe constipation which if not properly managed causes overflow pseudoincontinence and some have true fecal incontinence. For patients with anorectal malformations and those with idiopathic constipation with overflow pseudoincontinence, disimpaction, followed by an aggressive laxative regimen, often makes them continent, i.e. capable of having voluntary bowel movements. Surgical resection of a dilated rectosigmoid can dramatically reduce or eliminate these patients' daily laxative requirement, and improve their quality of life. For patients with true fecal incontinence, a bowel management program with a daily enema allows them to be kept artificially clean. For these patients, a surgical procedure whereby the daily enema is given antegrade is ideal, but it is the enema itself not its route which is the key to success. It is vital for the clinician to distinguish between these groups. Their initial presentations are quite similar, i.e. "fecal incontinence" or soiling but the treatments and surgical options differ dramatically. Much of what we have learned in the care of pediatric fecal incontinence can be extrapolated to adults.


Cell Cycle-independent Role of Cyclin D3 in Host Restriction of Influenza Virus Infection.
To identify new host factors that modulate the replication of influenza A virus, we performed a yeast two-hybrid screen using the cytoplasmic tail of matrix protein 2 from the highly pathogenic H5N1 strain. The screen revealed a high-score interaction with cyclin D3, a key regulator of cell cycle early G1 phase. M2-cyclin D3 interaction was validated through GST pull-down and recapitulated in influenza A/WSN/33-infected cells. Knockdown of Ccnd3 by small interfering RNA significantly enhanced virus progeny titers in cell culture supernatants. Interestingly, the increase in virus production was due to cyclin D3 deficiency per se and not merely a consequence of cell cycle deregulation. A combined knockdown of Ccnd3 and Rb1, which rescued cell cycle progression into S phase, failed to normalize virus production. Infection by influenza A virus triggered redistribution of cyclin D3 from the nucleus to the cytoplasm, followed by its proteasomal degradation. When overexpressed in HEK 293T cells, cyclin D3 impaired binding of M2 with M1, which is essential for proper assembly of progeny virions, lending further support to its role as a putative restriction factor. Our study describes the identification and characterization of cyclin D3 as a novel interactor of influenza A virus M2 protein. We hypothesize that competitive inhibition of M1-M2 interaction by cyclin D3 impairs infectious virion formation and results in attenuated virus production. In addition, we provide mechanistic insights into the dynamic interplay of influenza virus with the host cell cycle machinery during infection.


Correction of optical phase aberrations using binary-amplitude modulation.
We show that phase aberrations in an imaging system can be mitigated using binary-amplitude masks that reduce destructive interference in the image spatial frequency domain. Appropriately designed masks increase the magnitude of the optical transfer function and prevent nulls. This offers a low-cost, transmission-mode alternative to phase correction as used in active and adaptive optics, without a restriction on the waveband of operation.


Antibodies to basement membrane collagen and to laminin are present in sera from patients with poststreptococcal glomerulonephritis.
Sera from patients with poststreptococcal glomerulonephritis (PSGN) known to have antibodies to proteoglycans were studied for the presence of antibodies against other basement membrane (BM) components. BM collagen (type IV) was isolated in the native state by extracting bovine anterior lens capsule (ALC) with 0.5 M acetic acid. The 7-S (collagenous) domain and the NC-1 (noncollagenous) domain of type IV collagen were obtained after bacterial collagenase digestion of ALC followed by gel filtration. Laminin was isolated from the mouse EHS tumor and fibronectin from human plasma. Immunologic studies, using an ELISA and electroimmunoblot, revealed the presence of antibodies that reacted with intact, native type IV collagen and the 7-S collagenous domain of this molecule. Reaction with the NC-1 (noncollagenous) domain was minimal, and not higher than that obtained with control sera. Laminin reaction strongly with the patients' sera, but fibronectin did not. Unlike sera from patients with Goodpasture syndrome, which contain antibodies primarily against the NC-1 (noncollagenous) domain of type IV collagen, sera from patients with acute PSGN contain antibodies against all the major macromolecular components of BM. This difference in immunologic reactivity may account for the observed differences in the pathologic picture at the glomerular level.


Downregulation of Neuronal and Dendritic Connexin36-Made Electrical Synapses Without Glutamatergic Axon Terminals in Spinal Anterior Horn Cells From the Early Stage of Amyotrophic Lateral Sclerosis.
Connexin36 (Cx36) forms gap junctions between neurons, which are called electrical synapses, enabling adjacent neurons to communicate directly. The participation of chemical synapses in neurodegeneration in amyotrophic lateral sclerosis (ALS) has long been indicated, but it remains unclear whether electrical synapses are involved in the pathogenesis of ALS. We performed extensive immunopathological analyses using mutant superoxide dismutase 1 (SOD1G93A) transgenic mice and their littermates to investigate whether Cx36-made electrical synapses are affected in motor neuron diseases. We found that in the lamina IX of the lumbar spinal cord from wild type mice, about half of the Cx36 puncta existed independently of chemical synapse markers, while the rest coexisted with chemical synapse markers, such as vesicular glutamate transporter 1 (VGLUT1), which is a glutamatergic axon terminal marker, and/or glutamate decarboxylase 65 (GAD65), which is a GABAergic axon terminal marker. Cx36 single or Cx36/GAD65 double positive puncta, but not VGLUT1-containing puncta, were preferentially decreased on neuronal and dendritic surfaces of the anterior horn cells in the early stage of SOD1G93A ALS mice. Moreover, in five human autopsied sporadic ALS cases with bulbar or upper limb onset, Cx36 immunoreactivity was diminished in the proximal dendrites and neuropils of well-preserved large motor neurons in the lumbar anterior horns. These findings suggest that downregulation of neuronal and dendritic Cx36 in the spinal anterior horns commonly occurs from the early stage of hereditary and sporadic ALS. Cx36-made electrical synapses without glutamatergic signaling appear to be more vulnerable than other chemical synapses and electrical synapses with glutamatergic signaling in the early stage of motor neuron degeneration, suggesting involvement of Cx36-made electrical synapses in the pathogenesis of human ALS.


Mediolateral somitic origin of ribs and dermis determined by quail-chick chimeras.
Somites are transient mesodermal structures giving rise to all skeletal muscles of the body, the axial skeleton and the dermis of the back. Somites arise from successive segmentation of the presomitic mesoderm (PSM). They appear first as epithelial spheres that rapidly differentiate into a ventral mesenchyme, the sclerotome, and a dorsal epithelial dermomyotome. The sclerotome gives rise to vertebrae and ribs while the dermomyotome is the source of all skeletal muscles and the dorsal dermis. Quail-chick fate mapping and diI-labeling experiments have demonstrated that the epithelial somite can be further subdivided into a medial and a lateral moiety. These two subdomains are derived from different regions of the primitive streak and give rise to different sets of muscles. The lateral somitic cells migrate to form the musculature of the limbs and body wall, known as the hypaxial muscles, while the medial somite gives rise to the vertebrae and the associated epaxial muscles. The respective contribution of the medial and lateral somitic compartments to the other somitic derivatives, namely the dermis and the ribs has not been addressed and therefore remains unknown. We have created quail-chick chimeras of either the medial or lateral part of the PSM to examine the origin of the dorsal dermis and the ribs. We demonstrate that the whole dorsal dermis and the proximal ribs exclusively originates from the medial somitic compartment, whereas the distal ribs derive from the lateral compartment.


Acetaminophen interference with Nova StatStrip® Glucose Meter: case report with bench top confirmation.
Context: Point-of-care glucose meters are an integral part in the assessment of patients with altered mental status. For this reason, glucose meters are checked for interference from commonly encountered substances, including acetaminophen. The Nova StatStrip® glucose meter has previously been reported to be resistant to interference. We report a case of a very high acetaminophen concentration causing interference with this point-of-care glucose meter.Case report: A 25-year-old female presented after an overdose of acetaminophen and diphenhydramine combination product. Patient was minimally responsive, so a point-of-care glucose check was attempted using the Nova StatStrip® glucose meter. Five different meters were attempted, and each showed an error message. Laboratory analysis using Beckman Coulter® Unicel DxC 800 revealed a glucose of 180 mg/dL and an acetaminophen concentration of 465 mg/L. Serum spiked with acetaminophen at different concentrations revealed interference with the Nova StatStrip® glucose meter at a concentration of 399 mg/L and above. To our knowledge, this interference with the Nova StatStrip® glucose meter has not been reported in the medical literature.Conclusion: Very high levels of acetaminophen can interfere with point-of-care glucose meters, even those that have previously been reported to be robust such as the Nova StatStrip® glucose meter. Clinicians should be aware of this possible interference when treating patients with acetaminophen overdose.


Phosphorylation of the head domain of neurofilament protein (NF-M): a factor regulating topographic phosphorylation of NF-M tail domain KSP sites in neurons.
In neurons the phosphorylation of neurofilament (NF) proteins NF-M and NF-H is topographically regulated. Although kinases and NF subunits are synthesized in cell bodies, extensive phosphorylation of the KSP repeats in tail domains of NF-M and NF-H occurs primarily in axons. The nature of this regulation, however, is not understood. As obligate heteropolymers, NF assembly requires interactions between the core NF-L with NF-M or NF-H subunits, a process inhibited by NF head domain phosphorylation. Phosphorylation of head domains at protein kinase A (PKA)-specific sites seems to occur transiently in cell bodies after NF subunit synthesis. We have proposed that transient phosphorylation of head domains prevents NF assembly in the soma and inhibits tail domain phosphorylation; i.e. assembly and KSP phosphorylation in axons depends on prior dephosphorylation of head domain sites. Deregulation of this process leads to pathological accumulations of phosphorylated NFs in the soma as seen in some neurodegenerative disorders. To test this hypothesis, we studied the effect of PKA phosphorylation of the NF-M head domain on phosphorylation of tail domain KSP sites. In rat cortical neurons we showed that head domain phosphorylation of endogenous NF-M by forskolin-activated PKA inhibits NF-M tail domain phosphorylation. To demonstrate the site specificity of PKA phosphorylation and its effect on tail domain phosphorylation, we transfected NIH3T3 cells with NF-M mutated at PKA-specific head domain serine residues. Epidermal growth factor stimulation of cells with mutant NF-M in the presence of forskolin exhibited no inhibition of NF-tail domain phosphorylation compared with the wild type NF-M-transfected cells. This is consistent with our hypothesis that transient phosphorylation of NF-M head domains inhibits tail domain phosphorylation and suggests this as one of several mechanisms underlying topographic regulation.


Analysis of HOXB1 gene in a cohort of patients with sporadic ventricular septal defect.
Ventricular septal defect (VSD) including outlet VSD of double outlet right ventricle (DORV) and perimembranous VSD are among the most common congenital heart diseases found at birth. HOXB1 encodes a homeodomain transcription factor essential for normal cardiac outflow tract development. The aim of the present study was to investigate the possible genetic effect of sequence variations in HOXB1 on VSD. The coding regions and splice junctions of the HOXB1 gene were sequenced in 57 unrelated VSD patients. As a result, a homozygous c.74_82dup (p.Pro28delinsHisSerAlaPro) variant was identified in one individual with DORV. We also identified five previously reported polymorphisms (rs35114525, rs12946855, rs14534040, rs12939811, and rs7207109) in 18 patients (12 DORV and 6 perimembranous VSD). Our study did not show any pathogenic alterations in the coding region of HOXB1 among patients with VSD. To our knowledge this is the first study investigating the role of HOXB1 in nonsyndromic VSD, which provide more insight on the etiology of this disease.


First reported case of Cryptococcus gattii in the Southeastern USA: implications for travel-associated acquisition of an emerging pathogen.
In 2007, the first confirmed case of Cryptococcus gattii was reported in the state of North Carolina, USA. An otherwise healthy HIV negative male patient presented with a large upper thigh cryptococcoma in February, which was surgically removed and the patient was started on long-term high-dose fluconazole treatment. In May of 2007, the patient presented to the Duke University hospital emergency room with seizures. Magnetic resonance imaging revealed two large CNS lesions found to be cryptococcomas based on brain biopsy. Prior chest CT imaging had revealed small lung nodules indicating that C. gattii spores or desiccated yeast were likely inhaled into the lungs and dissemination occurred to both the leg and CNS. The patient's travel history included a visit throughout the San Francisco, CA region in September through October of 2006, consistent with acquisition during this time period. Cultures from both the leg and brain biopsies were subjected to analysis. Based on phenotypic and molecular methods, both isolates were C. gattii, VGI molecular type, and distinct from the Vancouver Island outbreak isolates. Based on multilocus sequence typing of coding and noncoding regions and virulence in a heterologous host model, the leg and brain isolates are identical, but the two differed in mating fertility. Two clinical isolates, one from a transplant recipient in San Francisco and the other from Australia, were identical to the North Carolina clinical isolate at all markers tested. Closely related isolates that differ at only one or a few noncoding markers are present in the Australian environment. Taken together, these findings support a model in which C. gattii VGI was transferred from Australia to California, possibly though an association with its common host plant E. camaldulensis, and the patient was exposed in San Francisco and returned to present with disease in North Carolina.


Performance assessment of a street-drainage bioretention system.
Event-based, flow-paced composite sampling was carried out at the inlet and outlet of a street-side bioretention facility in Seattle, Washington, to assess its ability to reduce street runoff quantity and pollutants. Over 2.5 years, 48 to 74% of the incoming runoff was lost to infiltration and evaporation. Outlet pollutant concentrations were significantly lower than those at the inlet for nearly all monitored constituents. In terms of mass, the system retained most of the incoming pollutants. Besides soluble reactive phosphorus (the mass of which possibly increased), dissolved copper was the least effectively retained; at least 58% of dissolved copper (and potentially as much as 79%) was captured by the system. Motor oil was removed most effectively, with 92 to 96% of the incoming motor oil not leaving the system. The results indicate that bioretention systems can achieve a high level of runoff retention and treatment in real-weather conditions.


Structural and functional similarities between the replication region of the Yersinia virulence plasmid and the RepFIIA replicons.
We sequenced the minimum replication region of the virulence plasmid pYVe439-80 from a serogroup O:9 Yersinia enterocolitica. This sequence is 68% homologous on a 1,873-nucleotide stretch to the sequence of the RepFIIA replicon of the resistance plasmid R100. The sequence contains two open reading frames, repA and repB, encoding proteins of 33,478 and 9,568 daltons, respectively. The amino acid sequences of the two proteins are 77 and 55% identical, respectively, to proteins RepA1 and RepA2 of the R100 replicon. Analysis of minicells transformed with a copy number mutant demonstrated that the replication region of pYVe439-80 directs the synthesis of a 33-kilodalton protein. Disruption of repA, encoding this protein, abolished replication. Two regions of pYVe439-80 are 76 and 70% homologous, respectively, to the copy number control antisense RNA and to the origin of replication region of R100. A mutation introduced in the pYVe439-80 DNA corresponding to the R100 sequence encoding the copy number control antisense RNA resulted in an increase in copy number, indicating a functional homology between the two replicons.


Tunable all-optical switching in periodic structures with liquid-crystal defects.
We suggest that tunable orientational nonlinearity of nematic liquid crystals can be employed for all-optical switching in periodic photonic structures with liquid-crystal defects. We consider a one-dimensional periodic structure of Si layers with a local defect created by infiltrating a liquid crystal into a pore, and demonstrate, by solving numerically a system of coupled nonlinear equations for the nematic director and the propagating electric field, that the light-induced Freedericksz transition can lead to a sharp switching and diode operation in the photonic devices.


Advanced In Vivo Prediction by Introducing Biphasic Dissolution Data into PBPK Models.
Coupling biorelevant in vitro dissolution with in silico physiological-based pharmacokinetic (PBPK) tools represents a promising method to describe and predict the in vivo performance of drug candidates in formulation development including non-passive transport, prodrug activation, and first-pass metabolism. The objective of the present study was to assess the predictability of human pharmacokinetics by using biphasic dissolution results obtained with the previously established BiPHa+ assay and PBPK tools. For six commercial drug products, formulated by different enabling technologies, the respective organic partitioning profiles were processed with two PBPK in silico modeling tools, namely PK-Sim and GastroPlus®, similar to extended-release dissolution profiles. Thus, a mechanistic dissolution/precipitation model of the assessed drug products was not required. The developed elimination/distribution models were used to simulate the pharmacokinetics of the evaluated drug products and compared with available human data. In essence, an in vitro to in vivo extrapolation (IVIVE) was successfully developed. Organic partitioning profiles obtained from the BiPHa+ dissolution analysis enabled highly accurate predictions of the pharmacokinetic behavior of the investigated drug products. In addition, PBPK models of (pro-)drugs with pronounced first-pass metabolism enabled adjustment of the solely passive diffusion predicting organic partitioning profiles, and increased prediction accuracy further.


New findings of the correlation between acupoints and corresponding brain cortices using functional MRI.
A preliminary study of the correlation between acupuncture points (acupoints) for the treatment of eye disorders suggested by ancient Oriental literature and the corresponding brain localization for vision described by Western medicine was performed by using functional MRI (fMRI). The vision-related acupoint (VA1) is located in the lateral aspect of the foot, and when acupuncture stimulation is performed there, activation of occipital lobes is seen by fMRI. Stimulation of the eye by directly using light results in similar activation in the occipital lobes by fMRI. The experiment was conducted by using conventional checkerboard 8-Hz light-flash stimulation of the eye and observation of the time-course data. This was followed by stimulation of the VA1 by using the same time-course paradigm as visual light stimulation. Results obtained with 12 volunteers yielded very clean data and very close correlations between visual and acupuncture stimulation. We have also stimulated nonacupoints 2 to 5 cm away from the vision-related acupoints on the foot as a control, and activation in the occipital lobes was not observed. The results obtained demonstrate the correlation between activation of specific areas of brain cortices and corresponding acupoint stimulation predicted by ancient acupuncture literature.


Evaluation of combined pathological responses in primary tumor and lymph nodes following neoadjuvant chemoimmunotherapy in non-small cell lung cancer.
Inconsistent pathological responses of tumor and lymph nodes (LNs) were frequently observed in non-small cell lung cancer (NSCLC) receiving neoadjuvant chemoimmunotherapy. However, there is a lack of studies to report the prognostic significance and the relevant clinicopathological factors of tumor-nodal inconsistent responses after neoadjuvant immunotherapy or chemoimmunotherapy. Therefore, this study aimed to depict the inconsistent pathological combined tumor-nodal responses in NSCLC patients after neoadjuvant chemoimmunotherapy as well as the underlying clinical significance. A total of 81 node-positive NSCLC patients who underwent neoadjuvant chemoimmunotherapy were eligible for inclusion. Demographic, radiologic, and pathological features of patients were recorded. Patients with pathological complete response of both tumor (ypT(pCR)) and LNs (ypN0) were classified into the combined good responder group and the relevant clinicopathological features were evaluated. The event-free survival (EFS) outcome was analyzed using Kaplan-Meier analysis. The ypN0 and ypT(pCR) rates were 74.1 % and 42.0 %, respectively. A significant correlation was observed between ypT(pCR) and ypN0 (P = 0.003), but inconsistent responses remained. The combined responses of the primary tumor and LNs demonstrated a significant association with the prognosis outcome (P = 0.005). Notably,patients who received at least twice of their infusions of immune checkpoint inhibitors after 15:30 had a worse prognosis (P = 0.015). A significant but not absolute correlation was observed between good tumor response and good nodal response in NSCLC patients after neoadjuvant chemoimmunotherapy, but inconsistent responses were also found. The combination of tumor and nodal responses is significantly associated with prognosis and combined good responder can be used as a reliable prognosis predictor.


Effects of Lower Extremity Muscle Fatigue on Knee Loading During a Forward Drop Jump to a Vertical Jump in Female Athletes.
The aim of this study was to examine changes in the kinematic and kinetic parameters of female athletes performing a forward drop jump to a vertical jump under muscle fatigue condition. Twelve female college athletes performed a forward drop jump to a vertical jump with and without muscle fatigue conditions. A motion capture system and two AMTI force plates were used to synchronously collect kinematic and kinetic data. Inverse dynamics were implemented to calculate the participant's joint loading, joint moment, and energy absorption. A paired sample t-test was used to compare statistical differences between pre-fatigue and post-fatigue conditions (α = .05). The forward trunk lean angle at initial foot contact, as well as the knee range of motion, total negative work and energy absorption contribution of the knee joint during the landing phase were significantly decreased under post-fatigue condition. The increased peak vertical ground reaction force and peak tibial anterior shear forces were also found under post-fatigue condition. These results indicated that muscle fatigue caused participants to change their original landing posture into stiff landing posture and decrease the energy absorption ability, which increased the tibial anterior shear forces. Therefore, female athletes should appropriately increase the knee flexion angle under muscle fatigue condition to reduce the risk of anterior cruciate ligament injuries.


Outcomes of physical activity for people living with dementia: qualitative study to inform a Core Outcome Set.
The need for a Core Outcome Set to evaluate physical activity interventions for people living with dementia, across stages of disease and intervention settings has been established. This qualitative study precedes the consensus phase of developing this Core Outcome Set and aims to: (i) compare the outcomes identified by patients, carers and professionals to those previously reported in the literature; (ii) and understand why certain outcomes are considered important. In-depth semi-structured interviews were conducted with people living with dementia, family carers and professionals (n=29). The outcomes identified in the interviews were mapped to a list of outcomes reported in a recent literature review. An in-depth thematic analysis was conducted to understand the importance of physical activity in dementia care. A comprehensive, inductively derived list of 77 outcomes, common across stages of dementia and intervention setting, was put together for the consensus phase of this Core Outcome Set: ten of these were new outcomes generated by this qualitative study. Five themes explained why stakeholders perceived physical activity outcomes as important for people living with dementia: "being well and staying well", "having a role towards others", "maintaining identity", "being connected to the present" and "delivering good quality care". Ten new outcomes of physical activity, not previously reported in recent literature, were identified. Physical activity is considered important to people living with dementia due to its positive impact on multiple health outcomes for both patients and carers.


Organotypic raft cultures as an effective in vitro tool for understanding Raman spectral analysis of tissue.
There is a growing body of evidence showing that optical spectroscopy has the potential to be a useful in vivo diagnostic tool. Yet, so far there is no definitive cellular and biochemical understanding for the differences seen in the spectra from different tissue categories and disease states. In this study, we examine the use of organotypic raft cultures as an in vitro model of in vivo tissue conditions in an attempt to overcome some of the limitations of previously used methods. Organotypic raft cultures resembling normal and dysplastic epithelial cervical tissue were constructed and grown at an air-liquid interface for 2 weeks. Raman spectra of normal as well as dysplastic raft cultures were measured and compared with in vivo spectra from the corresponding tissue type. Histologic comparisons ensured that the raft cultures had similar structure and morphology to the corresponding intact tissue types. Raman spectra were also acquired from different layers of tissue. Spectral comparisons show that the Raman spectra of the raft cultures are similar to the spectra acquired from the cervix in vivo for both normal and dysplastic tissues. These results show that organotypic raft cultures are an effective and useful tool for the cellular and biochemical analysis of tissue spectroscopy.


The influence of ortho-methyl substitution in organometallic self-assembly - a comparative study on Cu(111) vs. Ag(111).
Metal surface-induced dehalogenation of precursors is known to initiate self-assembly of organometallic networks, where tectons are connected via carbon-metal-carbon (C-M-C) bonds. Even though reversibility of the C-M-C bonds facilitates structural equilibration, defects associated with highly bent organometallic linkages are still commonly observed. By introducing a steric hindrance to reduce the C-M-C bond angle flexibility, we find well ordered organometallic networks of an ortho-methyl substituted 1,3,5-tris(p-bromophenyl)benzene analogue on Cu(111) after room-temperature (RT) deposition and on Ag(111) after annealing.


Glucan synthase complex of Aspergillus fumigatus.
The glucan synthase complex of the human pathogenic mold Aspergillus fumigatus has been investigated. The genes encoding the putative catalytic subunit Fks1p and four Rho proteins of A. fumigatus were cloned and sequenced. Sequence analysis showed that AfFks1p was a transmembrane protein very similar to other Fksp proteins in yeasts and in Aspergillus nidulans. Heterologous expression of the conserved internal hydrophilic domain of AfFks1p was achieved in Escherichia coli. Anti-Fks1p antibodies labeled the apex of the germ tube, as did aniline blue fluorochrome, which was specific for beta(1-3) glucans, showing that AfFks1p colocalized with the newly synthesized beta(1-3) glucans. AfRHO1, the most homologous gene to RHO1 of Saccharomyces cerevisiae, was studied for the first time in a filamentous fungus. AfRho proteins have GTP binding and hydrolysis consensus sequences identical to those of yeast Rho proteins and have a slightly modified geranylation site in AfRho1p and AfRho3p. Purification of the glucan synthase complex by product entrapment led to the enrichment of four proteins: Fks1p, Rho1p, a 100-kDa protein homologous to a membrane H(+)-ATPase, and a 160-kDa protein which was labeled by an anti-beta(1-3) glucan antibody and was homologous to ABC bacterial beta(1-2) glucan transporters.


Enhanced histamine-induced itch in diacylglycerol kinase iota knockout mice.
Subsets of small-diameter dorsal root ganglia (DRG) neurons detect pruritogenic (itch-causing) and algogenic (pain-causing) stimuli and can be activated or sensitized by chemical mediators. Many of these chemical mediators activate receptors that are coupled to lipid hydrolysis and diacylglycerol (DAG) production. Diacylglycerol kinase iota (DGKI) can phosphorylate DAG and is expressed at high levels in small-diameter mouse DRG neurons. Given the importance of these neurons in sensing pruritogenic and algogenic chemicals, we sought to determine if loss of DGKI impaired responses to itch- or pain-producing stimuli. Using male and female Dgki-knockout mice, we found that in vivo sensitivity to histamine-but not other pruritogens-was enhanced. In contrast, baseline pain sensitivity and pain sensitization following inflammatory or neuropathic injury were equivalent between wild type and Dgki-/- mice. In vitro calcium responses in DRG neurons to histamine was enhanced, while responses to algogenic ligands were unaffected by Dgki deletion. These data suggest Dgki regulates sensory neuron and behavioral responses to histamine, without affecting responses to other pruritogenic or algogenic agents.


Comparison of Antithrombin III and Pentoxifylline Treatments in Gram Negative Sepsis Patients Developing Disseminated Intravascular Coagulation.
The aim of this study was to evaluate the effects of antithrombin III (AT III) and pentoxifylline treatments on the gram negative septic patients with disseminated intravascular coagulation (DIC). For six days after plasma AT III activity dropped lower than 80% in Gram-patients who developed DIC were treated with AT III (90-120 IU/kg/day in 6 hours) or pentoxifylline (1.5 mg/kg/h in 6 hours) Fibrinogen, FDP, D-dimer, complete blood count, AT III activity, and DIC scores were calculated and recorded. The coagulation tests, AT III activity and FDP started to improve from the second day of treatment with both treatments (p<0.05). D-Dimer started to decrease on the second day of treatment with pentoxifylline (p<0.001) and fourth day of AT III treatment (p<0.05). Fibrinogen levels decreased on the second day of pentoxifylline treatment (p<0.05) and on the last day of AT III treatment (p<0.001). DIC scores started to decrease on the last day of treatment with AT III treatment (p<0.001) and on the third day of treatment with pentoxifylline (p<0.05). Both ATIII and pentoxifylline treatments had positive effects on fibrinogen, FDP, D-Dimer, AT III activity and DIC scores in patients with Gram-negative sepsis who developed DIC.


[Diagnosis of early extrauterine pregnancy by serum and Douglas pouch fluid hCG level].
In 44 patients with assumed ectopic pregnancy culdocentesis were carried out under transvaginosonographic control. In 23 cases, the clinical diagnosis ectopic pregnancy and in 16 cases, an early intact intrauterine pregnancy could be verified. Five patients had an early abortion. In all patients with an intact intrauterine pregnancy, the cul-de-sac liquid was serious. In 16 of the 23 cases of ectopic pregnancies, the fluid was bloody, 6 aspirates were bloody/serous and one serous. The findings in patients with an early abortion were as follows: three serous, one bloody/serous, one bloody. Further information could be gathered from the comparison of hCG-levels in simultaneously obtained serum samples and cul-de-sac liquid. All intact intrauterine pregnancies were associated with lower hCG-levels in the cul-de-sac liquid than in the serum (ratio: serum/cul-de-sac liquid greater than 1.3). In all ectopic pregnancies but one, we found an inverse constellation (ratio: serum/cul-de-sac liquid less than 0.7). In the described exceptional case the ratio was nearly 1 and when culdocentesis was repeated 5 days later, a ratio of 0.17 proved the clinically suspected ectopic pregnancy. In 3 cases of histologically confirmed intrauterine abortions, the hCG-level in the cul-de-sac liquid was lower than in the serum, in 2 cases of early abortions without histologically proved localisation the hCG-level was slightly above the level in the serum.


Improving detection and notification of tuberculosis cases in students in Shaanxi province, China: an intervention study.
Cooperation between different public and private health institutes involved in tuberculosis (TB) control has proven to enhance TB control in different settings. In China, such a mechanism has not been set up yet between Centers for Disease Control (CDCs) and university hospitals despite an increased TB incidence among students. This study aims to improve arrival of TB suspects identified by universities at the CDCs in order to manage them under standardized, directly observed treatment-short course (DOTS) conditions according to the National Tuberculosis Programme (NTP) guidelines. Five matched pairs of universities were randomly assigned to the control and intervention group. After a baseline survey, a cooperation mechanism between local CDCs and university hospitals was set up in the intervention group. The effects on referral of TB suspects to the local CDC, tracing by the local CDC, and arrival at the local CDCs were assessed. Differences were tested by means of the chi-square test. During the baseline survey, the referral, tracing and arrival rates were between 37% and 46%. After implementation of the cooperation mechanism, these rates had not changed in the control group but increased significantly in the intervention group: the referral, tracing and arrival rates were 97%, 95%, and 93%, respectively. It is feasible and effective to set up cooperation between CDCs and university hospitals to increase the number of TB suspects examined by CDCs and increase the number of TB patients treated under DOTS conditions. These public-public mix (PPM) activities should be expanded to cover all other university hospitals in China.


[Trends in medical education].
Trends in medical education include those occurring in medical practice, related with its contents, and in the educational sciences, related with methods and technics that could be employed. Trends in medical practice are related to epidemiologic, demographic and economic transitions, with the overwhelming influence of medical technology, specially molecular biology, the increasing social regulation, evidence based medicine and transdisciplinary contributions. In the field of pedagogy, trends include the acceptance of the strategic value of medical education, the importance of quality-and not only of covering, the attention to educational necessities, the recognition of the adulthood of most of the learners, the importance of its individual differences and the application of new educational technics.


Characterization of natural populations of Drosophila melanogaster with regard to the hobo system: a new hypothesis on the invasion.
Until now, with regard to the hobo system of hybrid dysgenesis, natural populations of Drosophila melanogaster have been investigated using only two criteria: at the molecular level, the presence or absence of XhoI fragments 2.6 kb long or smaller; and/or at the genetic level, the ability to induce gonadal dysgenesis sterility in crosses A (females of an E reference strain crossed with males under test) and A* (females under test crossed with males of an H reference strain). Recently, analyses of laboratory strains using these criteria as well as the mobilization of two reporter genes, the male recombination and the number of 'TPE' repeats in the S region, revealed a lack of correlation between the different dysgenic parameters themselves, and also between these parameters and the molecular characteristics of the strains. Thirteen current strains derived from world populations were therefore investigated with regard to all these dysgenic traits, to determine discriminating criteria providing a robust method of classifying natural populations and deducing the dynamics of hobo elements in these populations. We show, as in laboratory strains, a lack of correlation between the parameters studied. Therefore, the significance of each of them as well as the nature of hobo hybrid dysgenesis are discussed, to propose an analysis method of the hobo system applicable to natural populations. According to the geographical distribution of hobo activities in world populations and to the variable polymorphism of the number of 'TPE' repeats, we propose a new scenario for the invasion of D. melanogaster by hobo elements.


Electrically driven directional motion of a four-wheeled molecule on a metal surface.
Propelling single molecules in a controlled manner along an unmodified surface remains extremely challenging because it requires molecules that can use light, chemical or electrical energy to modulate their interaction with the surface in a way that generates motion. Nature's motor proteins have mastered the art of converting conformational changes into directed motion, and have inspired the design of artificial systems such as DNA walkers and light- and redox-driven molecular motors. But although controlled movement of single molecules along a surface has been reported, the molecules in these examples act as passive elements that either diffuse along a preferential direction with equal probability for forward and backward movement or are dragged by an STM tip. Here we present a molecule with four functional units--our previously reported rotary motors--that undergo continuous and defined conformational changes upon sequential electronic and vibrational excitation. Scanning tunnelling microscopy confirms that activation of the conformational changes of the rotors through inelastic electron tunnelling propels the molecule unidirectionally across a Cu(111) surface. The system can be adapted to follow either linear or random surface trajectories or to remain stationary, by tuning the chirality of the individual motor units. Our design provides a starting point for the exploration of more sophisticated molecular mechanical systems with directionally controlled motion.


Decision making in laboratory test studies.
Evaluation of laboratory tests can be made at multiple levels. Evaluation at the levels of operational and decision making usefulness may be the most important but most neglected areas of analysis. Although quantitative models for such analysis are available, their application is often hindered by lack of precise probability measurements, unavailability of sensitivity and specificity data, and a poor understanding of the decision making process by physicians. A general understanding of such models and their mathematic principles have been presented with the hope that this will improve the diagnostic usefulness and application of the laboratory tests to be discussed in the remainder of this volume.


[Experimental characteristics of anthrax intoxication].
In experimental anthrax intoxication, a highly important stage of its pathogenesis consists in microcirculatory disturbances with the phenomena of blood sludge, accompanied by the increased permeability of blood vessels not only for plasma, but also for red blood cells. These disturbances result in perivascular hemorrhages, hemorrhagic infiltrations, edema and cavitary transudates. Pulmonary edema and, as a consequence, the accumulation of fluid in pulmonary alveoli and the respiratory tract are of particular importance and, probably, can be considered the basic cause of the ensuing acute and fatal asphyxia. Such vascular and pulmonary insufficiency is accompanied by a decrease in the content of macroergic compounds (and in particular ATP), the characteristic deformation of red blood cells and disturbances in their oxygen transport function, which is linked with the decreased content of 2,3-diphosphoglycerate.


Long-term outcome after callosotomy or vagus nerve stimulation in consecutive prospective cohorts of children with Lennox-Gastaut or Lennox-like syndrome and non-specific MRI findings.
There is currently no resective (potentially curative) surgical option that is useful in patients with Lennox-Gastaut syndrome. Palliative procedures such as callosotomy (Cx), vagus nerve stimulation (VNS) or deep brain stimulation have been offered. We compared the outcomes after Cx or VNS in two consecutive prospective cohorts of patients with generalised epilepsy. Twenty-four patients underwent callosotomy from 2006 to 2007 (Group 1); 20 additional patients were submitted to VNS from 2008 to 2009 (Group 2). They had generalised epilepsy of the Lennox-Gastaut or Lennox-like type. They were submitted to a neurological interview and examination, interictal and ictal video-EEG, high resolution 1.5T MRI, and cognitive and quality of life evaluations. The two-year post-operative follow-up results were evaluated for each patient. The final mean stimuli intensity was 3.0 mA in the Group 2 patients. Seizure-free patients accounted for 10% in Group 1 and none in Group 2. Ten and sixteen percent of the Group 1 and 2 patients, respectively, were non-responders. Improvements in attention and quality of life were noted in 85% of both Group 1 and 2 patients. Rupture of the secondary bilateral synchrony was noted in 85% of Group 1 patients; there was no EEG modification after VNS in Group 2. Both procedures were effective regarding the control of atypical absences and generalised tonic-clonic seizures. Both procedures were not effective in controlling tonic seizures. Callosotomy was very effective in reducing the frequency of atonic seizures, but VNS was ineffective. In contrast, callosotomy was not effective in reducing myoclonic seizures, whereas VNS was. Callosotomy might be preferred as the primary treatment in children with Lennox-Gastaut syndrome, and no specific findings on MRI if atonic seizures prevail in the patient's clinical picture; when myoclonic seizures prevail, the same might hold true in favour of VNS. When atypical absence or generalised tonic-clonic seizures are the main concern, although both procedures carry similar effectiveness, VNS might be considered a good option as an initial approach, taking into account the adverse event profile. Patients should be advised that both procedures are not very effective in the treatment of tonic seizures.


Beneficial effects of autologous mesenchymal stem cell transplantation in active progressive multiple sclerosis.
In this study (trial registration: NCT02166021), we aimed to evaluate the optimal way of administration, the safety and the clinical efficacy of mesenchymal stem cell (MSC) transplantation in patients with active and progressive multiple sclerosis. Forty-eight patients (28 males and 20 females) with progressive multiple sclerosis (Expanded Disability Status Scale: 3.0-6.5, mean : 5.6 ± 0.8, mean age: 47.5 ± 12.3) and evidence of either clinical worsening or activity during the previous year, were enrolled (between 2015 and 2018). Patients were randomized into three groups and treated intrathecally (IT) or intravenously (IV) with autologous MSCs (1 × 106/kg) or sham injections. After 6 months, half of the patients from the MSC-IT and MSC-IV groups were retreated with MSCs, and the other half with sham injections. Patients initially assigned to sham treatment were divided into two subgroups and treated with either MSC-IT or MSC-IV. The study duration was 14 months. No serious treatment-related safety issues were detected. Significantly fewer patients experienced treatment failure in the MSC-IT and MSC-IV groups compared with those in the sham-treated group (6.7%, 9.7%, and 41.9%, respectively, P = 0.0003 and P = 0.0008). During the 1-year follow-up, 58.6% and 40.6% of patients treated with MSC-IT and MSC-IV, respectively, exhibited no evidence of disease activity compared with 9.7% in the sham-treated group (P < 0.0001 and P < 0.0048, respectively). MSC-IT transplantation induced additional benefits on the relapse rate, on the monthly changes of the T2 lesion load on MRI, and on the timed 25-foot walking test, 9-hole peg test, optical coherence tomography, functional MRI and cognitive tests. Treatment with MSCs was well-tolerated in progressive multiple sclerosis and induced short-term beneficial effects regarding the primary end points, especially in the patients with active disease. The intrathecal administration was more efficacious than the intravenous in several parameters of the disease. A phase III trial is warranted to confirm these findings.


Vasoactive intestinal peptide: a possible transmitter of nonadrenergic relaxation of guinea pig airways.
Vasoactive intestinal peptide, a smooth-muscle relaxant neuropeptide with neurotransmitter properties, was relaxed during electrical field stimulation of guinea pig trachea. The amount released correlated with the degree of relaxation, and the release was blocked by tetrodotoxin. Prior incubation of the trachea with antiserum to vasoactive intestinal peptide reduced the relaxation. Thus vasoactive intestinal peptide may mediate the nonadrenergic relaxation of tracheal smooth muscle.


Long-term safety and efficacy of pegunigalsidase alfa: A multicenter 6-year study in adult patients with Fabry disease.
Fabry disease (FD) is a rare lysosomal storage disorder caused by pathogenic variants in the GLA gene encoding α-galactosidase (α-Gal)-A. We evaluated long-term safety/efficacy of pegunigalsidase alfa, a novel PEGylated α-Gal-A enzyme replacement therapy (ERT) now approved for FD. In a phase-1/2 dose-ranging study, 15 ERT-naive adults with FD completed 12 months of pegunigalsidase alfa and enrolled in this 60-month open-label extension of 1 mg/kg pegunigalsidase alfa infusions every 2 weeks. Fifteen patients enrolled (8 males; 7 females); 10 completed ≥48 months (60 months total treatment), and 2 completed 60 months (72 months total treatment). During treatment, most treatment-emergent adverse events were mild/moderate in severity and all infusion-related reactions were mild/moderate in severity. Four patients were transiently positive for anti-pegunigalsidase alfa IgG. Patients showed continuous reduction in plasma lyso-Gb3 concentrations with mean (standard error) reduction of 76.1 [25.1] ng/mL from baseline to month 24. At 60 months, the estimated glomerular filtration rate slope was comparable to that observed in patients treated with other ERTs. Cardiac function assessments revealed stability; no cardiac fibrosis was observed. In this first long-term assessment of pegunigalsidase alfa administration in patients with FD, we found favorable safety/efficacy. Our data suggest long-term continuous benefits of pegunigalsidase alfa treatment in adults with FD.


Human myeloma cell line carrying a Philadelphia chromosome.
A new human plasmacytoma cell line (Karpas 707) has been established from a myeloma patient. The cultured cells are negative for Epstein-Barr viral nuclear antigen and free of mycoplasma. They are similar to plasma cells and secrete only lambda light chains. The cells are hypodiploid and contain the Philadelphia chromosome and other abnormalities. This cell line may be suitable for the production of human monoclonal antibodies.


Activation of NK cell cytotoxicity by the natural compound 2,3-butanediol.
The natural compound 2,3-BTD has diverse physiological effects in a range of organisms, including acting as a detoxifying product of liver alcohol metabolism in humans and ameliorating endotoxin-induced acute lung injury in rats. In this study, we reveal that 2,3-BTD enhances NK cell cytotoxic activity in human pNK cells and NK92 cells. Treatment of NK cells with 2,3-BTD increased perforin expression in a dose-dependent manner. This was accompanied by elevated JNK and ERK1/2 MAPK activities and enhanced expression of NKG2D/NCRs, upstream signaling molecules of the MAPK pathways. The 2,3-BTD effect was inhibited by pretreatment with inhibitors of JNK (SP) or ERK1/2 (PD) or by depleting NKG2D/NCRs or JNK1 or ERK2 with siRNA. These results indicate that 2,3-BTD activates NK cell cytotoxicity by NKG2D/NCR pathways and represent the first report of the 2,3-BTD effect on activation of innate immunity cells.


[Modern systemic therapy of advanced pancreatic cancer: impact on overall survival and medico economical aspects].
More than 75% of patients with advanced pancreas cancer present at a later stage, when only palliative chemotherapy can be offered as the main treatment. Since introduced, gemcitabine has significantly improved the symptomatic impact of the disease, and this single-agent therapy still remains the gold-standard treatment aside of any exploratory protocol. Survival benefits have however been marginal, although a minority of long-term survivors now survives their disease up to two or three years. Results from a retrospective study conducted on cases followed by the oncosurgery service of the Geneva University Hospital from 1993 to 2003 support this observation, and show a trend towards a parallel increase in ambulatory-incurred costs.


The case of the missing ring: radical cleavage of a carbon-carbon bond and implications for GFP chromophore biosynthesis.
The green fluorescent protein (GFP) creates its fluorophore by promoting spontaneous peptide backbone cyclization and amino acid oxidation chemistry on its own Ser65, Tyr66, Gly67 tripeptide sequence. Here we use high-resolution crystallography and mutational analyses to characterize GFP variants that undergo backbone cyclization followed by either anticipated chromophore synthesis via Y66F Calpha-Cbeta double-bond formation or unprecedented loss of a Y66F benzyl moiety via Calpha-Cbeta bond cleavage. We discovered a Y66F cleavage variant that subsequently incorporates an oxygen atom, likely from molecular oxygen, at the Y66 Calpha position. The post-translational products identified from these Y66F GFP structures support a common intermediate that partitions between Calpha-Cbeta oxidation and homolytic cleavage pathways. Our data indicate that Glu222 is the branchpoint control for this partitioning step and also influences subsequent oxygen incorporation reactions. From these results, we propose mechanisms for Y66F Calpha-Cbeta cleavage, oxygen incorporation, and chromophore biosynthesis with shared features that include radical chemistry. By revealing how GFP and RFP protein environments steer chemistry to favor fluorophore biosynthesis and disfavor alternative reactivity, we identify strategies for protein design. The proposed, common, one-electron oxidized, radical intermediate for post-translation modifications in the GFP family has general implications for how proteins drive and control spontaneous post-translational chemical modifications in the absence of metal ions.


National and Provincial Prevalence of Pediculus humanus capitis among Urban Students in Iran from 2014 to 2018.
Pediculosis capitis is one of the most common contagious diseases in overcrowded places, especially in schools. We aimed to determine the overall and seasonal prevalence of Pediculosis capitis (head lice infestation) among Iranian students. A repeated cross-sectional study. The present study was conducted among all students, aged 6-18 yr, from urban areas of 31 provinces of Iran from 2014 to 2018. The student's hair and scalp were examined by trained school health instructor. The prevalence (95% Confidence Interval) of pediculosis were estimated overall and stratified by gender. Linear and Poisson regression models were used for data analysis. Results showed an upward trend of pediculosis from 1.7% to 3.42% during four years of study (P for trend=0.006). Overall, the prevalence of this infestation was significantly higher in girls than boys (P-value<0.05). The relative risk of head lice infestation in autumn was 2.42 times higher than spring (RR: 2.42, 95%CI: 1.35 to 4.32). Pediculosis is still a health issue among Iranian students, especially in girls. This infestation had upward trend in recent years and was more common in southern Iran. Increasing the awareness of students regarding symptoms and complications of pediculosis and using accurate diagnosis methods can be effective in reducing the prevalence of pediculosis and its consequences.


Relationship between genetic polymorphism of glutathione S-transferase-p1 and p53 protein accumulation in Iranian esophageal squamous cell carcinoma patients.
It has been reported that the activity of glutathione S-transferase (GST) is over-expressed in plasma and esophagus biopsies in Iranian patients suffering from esophageal squamous cell carcinoma (SCC). The aim of this study was to find out the frequency of GST-P genotypes in these patients. Moreover, the association of GST-P genotypes with p53 protein accumulation in esophageal epithelium was investigated. DNA isolated from paraffin-embedded tissue biopsies from patients suffering from esophageal SCC (n = 56) were collected. polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) using Alw261 enzyme was applied to determine GST-P genotypes (Ile 105 Val). All the samples were also subjected to immunohistochemistry (IHC) for p53. The frequency of GST-P genotypes in Iranian esophagus SCC patients for Ile/Ile, Ile/Val and Val/Val was 73.2, 21.5 and 5.3%. There was no association between GST-P polymorphism and p53 accumulation in esophageal epithelial cells. The frequency of GST-P polymorphism was not associated with p53 protein accumulation in esophagus epithelium. The frequency of polymorphic variants of GST-P, Ile/Ile, Ile/Val and Val/Val in SCC patients may suggest that Ile to Val substitution in GST-P gene dose not represent susceptibility to SCC in high-risk Iranian population.


A population-based study of the usefulness of screening for neuroblastoma.
Neuroblastoma has many characteristics which suggest that preclinical detection might improve outcome. The Quebec Neuroblastoma Screening Project was initiated to determine whether mass screening could reduce mortality in a large cohort of infants. As an early endpoint, we report whether screening could reduce the incidence of poor-prognosis neuroblastoma in children with advanced-stage disease over 1 year of age. All 476,603 children born in the province of Quebec during the 5-year period of May 1, 1989, to April 30, 1994, were eligible for urinary assay of homovanillic acid and vanillylmandelic acid at 3 weeks and 6 months of age. Children with a positive screen were referred to one of four paediatric cancer centres in the province for uniform evaluation and treatment if necessary. Standardised incidence ratios (SIRs) were calculated for neuroblastoma in the province and two similar population-based controls, the state of Minnesota and the province of Ontario, during the same period of time and with similar ascertainment procedures. Compliance with screening in Quebec province was 91% at 3 weeks (n = 425,816) and 74% at 6 months (n = 349,706). Through July 31, 1995, with a follow-up of the birth cohort of 15-75 months, 118 cases of neuroblastoma were diagnosed, 43 detected preclinically by screening, 20 detected clinically before screening at 3 weeks of age, and 55 detected clinically after 3 weeks of age having normal screens (52) or never screened (3). Retrospective analysis of stored samples confirmed that 49 of 52 patients missed by screening had levels of catecholamine metabolites that were too low to be detected at 6 months or earlier. Based on US Surveillance, Epidemiology and End Results data, 54.5 cases of neuroblastoma would have been expected in Quebec province during the study period, for an SIR of 2.17 (95% CI 1.79-2.57, p < 0.0001). For the two control groups, 43 and 80 cases of neuroblastoma were detected, respectively, compared with 37.9 and 85.4 expected, overall SIR 1.00 (not significant). SIRs for Quebec province by age at diagnosis in yearly intervals show a marked increased incidence under 1 year of age (SIR 2.85, 2.26-3.50), with no reduction in incidence in subsequent years. Limiting analysis to only patients diagnosed over 1 year of age with advanced-stage disease, 22 cases were detected in Quebec province versus 14.4 expected (SIR 1.52, 0.95-2.23). Data in the two control groups show no significant increase or decrease in any-stage disease in children under or over the age of 1 year, except for an increase in early-stage disease in Minnesota children over 1 year: 10 versus 3.8 expected (SIR 2.67, 1.27-4.58). Screening for neuroblastoma increases the incidence in infants without decreasing the incidence of unfavourable advanced-stage disease in older children. It is unlikely that screening for neuroblastoma in infants will reduce mortality for this disease.


Development and testing of acoustically-matched hydrogel-based electrodes for simultaneous EMG-ultrasound detection.
In this manuscript we describe the development and testing of a bipolar electrode for the simultaneous acquisition of ultrasound (US) images and surface electromyograms (EMGs) from the same muscle region. The developed electrode (bEMG-US) consists of two circular sensing regions (20 mm diameter) with fixed inter-electrode distance (3.5 cm, center-to-center). Both the sensing regions and the external structure of the electrode are made of hydrogel layers separated by insulating materials. The electrical properties (i.e., impedance and noise of the electrode-skin interface) and the quality of EMGs detected with the developed electrodes during electrically elicited contractions were assessed and compared with those provided by commercially available EMG electrodes. The effect of the bEMG-US electrode on US images was evaluated by comparing images detected from the same muscle region with and without the electrode interposed between the US probe and the skin. Tests on five subjects showed that the electrode-skin impedance of bEMG-US electrodes was higher than that of conventional EMG electrodes (mean (range): 15.6 (8.5-21.1) kΩ vs. 8.2 (4.9-16.5) kΩ). Despite higher impedance values, both electrode systems provided comparable, electrode-skin noise levels (1.4 (1.1-1.7) µV vs. 1.3 (1.0-1.5) µV) and M waves (normalized mean square error: 2.6 (0.6-6.8)%). The quality of US images detected with and without the bEMG-US electrode between the US probe and the skin was comparable, as demonstrated by the low errors in the estimation of anatomical variables in the two experimental conditions (range: (0.37-2.35) deg for pennation angle and (-0.31-0.1) cm for muscle thickness). Results demonstrate that bEMG-US can be used to acquire concurrently EMGs and US images from the same muscle region with a negligible effect on the quality of the two detected signals, thus allowing for a simultaneous, multimodal evaluation of muscle activation.


Interactions of the thymic polypeptide hormone thymopoietin with neuronal nicotinic alpha-bungarotoxin binding sites and with muscle-type, but not ganglia-type, nicotinic acetylcholine receptor ligand-gated ion channels.
Studies were conducted to assess the ability of the thymic polypeptide hormone thymopoietin (TPO) to interact with proto-typical ganglia-type or muscle-type nicotinic acetylcholine receptor ion channels (nAcChoR). Also investigated were interactions of TPO with neuronal nicotinic alpha-bungarotoxin binding sites (nBgtS), which share many features with nAcChoR and may belong to an extended nAcChoR family but do not appear to function as simple ligand-gated ion channels. TPO and alpha-bungarotoxin (Bgt) share the capacity for high affinity (IC50 values in the nanomolar range) interaction with nBgtS, which are expressed as high affinity radioiodinated Bgt binding sites by cells of the SH-SY5Y or IMR-32 human neuroblastomas. TPO and Bgt also share the capacity for high affinity interaction with muscle-type nAcChoR, which are expressed as high affinity binding sites for radioiodinated Bgt or tritium-labeled acetylcholine by cells of the TE671/RD human clone or the BC3H-1 mouse muscle cell line or on membrane preparations from Torpedo electroplax. TPO and Bgt act acutely as high affinity antagonists of muscle-type nAcChoR functional responses, which are measured using an isotopic rubidium ion efflux assay, on TE671/RD or BC3H-1 cells. In contrast, neither TPO nor Bgt are effective, at doses of up to 1 microM, as antagonists of ganglia-type nAcChoR function on SH-SY5Y or IMR-32 cells, nor are they potent as inhibitors of high affinity tritium-labeled acetylcholine binding to sites on putative ganglia-type nAcChoR expressed by SH-SY5Y or IMR-32 cells. These data indicate that some members of the extended nAcChoR family, including nBgtS and functional muscle-type nAcChoR but not ganglia-type nAcChoR, can interact with either Bgt or TPO. The results suggest that TPO may be an endogenous ligand active in both the nervous and immune systems and that some of its actions may be mediated via nBgtS or via functional blockade of muscle-type nAcChoR.


Overall and abdominal adiposity in midlife and subsequent cognitive function.
The scientific literature supports a link between midlife adiposity and cognitive function or decline but most studies to-date have investigated only overall adiposity, often omitting important confounders from the analyses. We investigated in a cross-time design the relationships between two different midlife adiposity markers and subsequent cognitive function, testing midlife dietary patterns as a potential confounder of the associations. The study was based on the «Supplémentation en Vitamines et Minéraux Antioxydants» randomized trial (SU.VI.MAX, 1994-2002) and the SU.VI.MAX 2 observational follow-up study (2007-2009). A general-population cohort in France. N=2,817 individuals (1,493 men and 1,324 women) included in both the SU.VI.MAX and SU.VI.MAX 2 studies. The cognitive performance of 2,817 middle-aged adults participating in the SU.VI.MAX (Supplémentation en Vitamines et Minéraux Antioxydant) study was assessed in 2007-2009 using 6 neuropsychological tests. Principal component analysis was used to derive specific cognitive scores. A composite cognitive score was also computed. Body mass index (BMI) and waist circumference (WC) were measured 13 years earlier (1994). Associations between midlife adiposity and cognitive functioning were estimated through covariance analyses. After adjustment for obesity-related cardio-metabolic parameters, higher BMI and larger WC at midlife predicted lower executive function. For example, the adjusted mean difference (95% confidence interval) for 1 SD increase in WC was -0.48 (-0.97, 0.00). Obese participants in midlife showed an adjusted mean difference (95% confidence interval) of -1.68 (-3.15, -0.22) compared with non-obese. Further adjustment for midlife dietary patterns slightly attenuated these associations. No relationships were observed with verbal memory or global cognitive function. Midlife overall and abdominal adiposity were similarly associated with lower executive functioning scores. Dietary patterns may partly explain such a relationship, arguing for the importance of controlling for lifestyle confounders in future studies.


Protection of European domestic pigs from virulent African isolates of African swine fever virus by experimental immunisation.
African swine fever (ASF) is an acute haemorrhagic disease of domestic pigs for which there is currently no vaccine. We showed that experimental immunisation of pigs with the non-virulent OURT88/3 genotype I isolate from Portugal followed by the closely related virulent OURT88/1 genotype I isolate could confer protection against challenge with virulent isolates from Africa including the genotype I Benin 97/1 isolate and genotype X Uganda 1965 isolate. This immunisation strategy protected most pigs challenged with either Benin or Uganda from both disease and viraemia. Cross-protection was correlated with the ability of different ASFV isolates to stimulate immune lymphocytes from the OURT88/3 and OURT88/1 immunised pigs.


The prediction of acute toxicity (LD50) for organophosphorus-based chemical warfare agents (V-series) using toxicology in silico methods.
Nerve agents are organophosphate chemical warfare agents that exert their toxic effects by irreversibly inhibiting acetylcholinesterase, affecting the breakdown of the neurotransmitter acetylcholine in the synaptic cleft. Due to the risk of exposure to dangerous nerve agents and for animal welfare reasons, in silico methods have been used to assess acute toxicity safely. The next-generation risk assessment (NGRA) is a new approach for predicting toxicological parameters that can meet modern requirements for toxicological research. The present study explains the acute toxicity of the examined V-series nerve agents (n = 9) using QSAR models. Toxicity Estimation Software Tool (ver. 4.2.1 and ver. 5.1.2), QSAR Toolbox (ver. 4.6), and ProTox-II browser application were used to predict the median lethal dose. The Simplified Molecular Input Line Entry Specification (SMILES) was the input data source. The results indicate that the most deadly V-agents were VX and VM, followed by structural VX analogues: RVX and CVX. The least toxic turned out to be V-sub x and Substance 100A. In silico methods for predicting various parameters are crucial for filling data gaps ahead of experimental research and preparing for the upcoming use of nerve agents.


[Validity of attentional-style subscales for the Japanese version of the Test of Attentional and Interpersonal Style (TAIS)].
The six attentional-style subscales of the Test of Attentional and Interpersonal Style (TAIS, Nideffer, 1976) have been used mainly in the sport psychology area to investigate the relationship between attentional abilities and performance. This study assessed the validity of the Japanese version of the TAIS attentional-style subscales with 157 male and 163 female students. A confirmatory factor analysis failed to validate Nideffer's attentional dimension of bandwidth (narrow to broad attention) and direction (internal to external). The findings replicate previous studies which showed that the structure of the TAIS attention-related subscales is not reasonable and its factorial validity does not meet the needs of psychometrics.


Metabolic characterization of the genus Brucella. III. Oxidative metabolism of strains that show anomalous characteristics by conventional determinative methods.
Meyer, Margaret E. (University of California, Davis). Metabolic characterization of the genus Brucella. III. Oxidative metabolism of strains that show anomalous characteristics by conventional determinative methods. J. Bacteriol. 82:401-410. 1961.-The oxidative metabolic patterns were determined on 83 strains of brucellae that had been described as "atypical" because they differed in one or more characteristics or because they had been isolated from an abnormal host (other than the natural reservoir for that species). Of the 83 strains examined, 44 displayed the metabolic pattern for Brucella melitensis. A comparison was then made between the results of identifying these strains metabolically and by the conventional methods. It was found that a few strains of B. melitensis showed a decreased tolerance to basic fuchsin and thionin, but none of the strains that was identified metabolically as B. melitensis produced H(2)S or required CO(2). No biotypes have been reported for this species, since only slight quantitative variation in dye tolerances occurs among strains of B. melitensis, and no metabolic variants were found. It is concluded that B. melitensis is a homogenous species and can be identified with certainty by its oxidative metabolic pattern, irrespective of its host or geographic source. Of the remaining strains, 38 displayed the metabolic pattern singular for Brucella abortus. Evidence was presented to support the conclusion that in this species the characteristics of dye tolerance, H(2)S production, and CO(2) required for initial growth vary independently of each other, and strains that differ from the species description by these criteria can be identified correctly by their oxidative metabolic pattern. Of the 83 atypical strains examined, 24 were strains of Brucella described as a new species, Brucella intermedia (Renoux). Of these 24 strains, 10 were identified as Brucella melitensis, 13 as Brucella abortus, and one as Brucella suis. Evidence was presented to support the conclusion that B. intermedia (Renoux) cannot be considered as a new species.


Mechanical support for acutely failed heart or lung grafts.
Heart and lung allograft dysfunction continues to be a problem in thoracic transplantation. Although medical therapy is often sufficient to restore allograft function, occasionally more invasive means are required. Mechanical assist devices, inhaled nitric oxide (iNO), and extracorporeal membrane oxygenation (ECMO) have been used with a modest degree of success in cases of refractory heart, lung, and heart-lung allograft failure. Allograft failure secondary to pulmonary hypertension often responds to iNO concentrations between 5 and 70 ppm without major toxicity. More severe cases may require mechanical assist devices or ECMO and carry higher risks of complications such as bleeding, neurological injury, and death. Utilization of and weaning from these interventions require intensive monitoring. Randomized, prospective studies are not ethically feasible, but case reports and patient series indicate the usefulness of mechanical circulatory support, iNO, and ECMO. This review focuses on the indications, complications, and patient survival rates associated with these modalities.


[SPECT and PET in neurobiology].
PET (positron emission tomography) and SPECT (single photon emission computed tomography) are isotopic methods in which the distribution is registered of radiolabelled tracers given in such small amounts that they are without effect on the organism or the organism's disposal of them. Thus, a series of important biological processes in the intact organism can be studied. The methods have been used in many disciplines but in particular for neurobiological research on the brain--e.g., the brain's regional blood circulation and mapping of the brain's functional structure. The methods have also been used in the investigation of glucose and amino acid metabolism in the brain and receptor conditions.


Agaricus blazei Murill Polysaccharides Protect Against Cadmium-Induced Oxidative Stress and Inflammatory Damage in Chicken Spleens.
Agaricus blazei Murill polysaccharide (ABP) has exhibited antioxidant and immunoregulatory activity. The aim of this study was to investigate the effect of ABP on cadmium (Cd)-induced antioxidant functions and inflammatory damage in chicken spleens. In this study, groups of 7-day-old chickens were fed with normal saline (0.2 mL single/day), CdCl2 (140 mg/kg/day), ABP (30 mg/mL, 0.2 mL single/day), and Cd + ABP (140 mg/kg/day + 0.2 mL ABP). Spleens were separated on the 20th, 40th, and 60th day for each group. The Cd contents, expression of melanoma-associated differentiation gene 5 (MDA5) and its downstream signaling molecules (interferon promoter-stimulating factor 1 (IPS-1), transcription factors interferon regulatory factor 3 (IRF3), and nuclear factor kappa-light chain-enhancer of activated B cells (NF-κB)), the content of cytokines (interleukin 1β (IL-1β), interleukin 6 (IL-6), tumor necrosis factor-α (TNF-α) and beta interferon (IFN-β)), protein levels of heat shock proteins (HSPs), levels of malondialdehyde (MDA), activities of glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD), and histopathological changes of spleens were detected on the 20th, 40th, and 60th day. The results showed that ABP significantly reduced the accumulation of Cd in the chicken spleens and reduced the expression of MDA5, IPS-1, IRF-3, and NF-κB; their downstream inflammatory cytokines, IL-1β, IL-6, TNF-α, and IFN-β; and the protein levels of HSPs (HSP60, HSP70, and HSP90) in spleens. The activities of antioxidant enzymes (SOD and GSH-Px) significantly increased, and the level of MDA decreased in the ABP + Cd group. The results indicate that ABP has a protective effect on Cd-induced damage in chicken spleens.


Surgical management of 121 benign proximal fibula tumors.
Tumors of the fibula comprise only 2.5% of primary bone lesions. Patients with aggressive benign tumors in the proximal fibula may require en bloc resection. Peroneal nerve function, knee stability, and recurrence are substantial concerns with these resections. The incidence and fate of these complications is not well-known owing to the small numbers of patients in previous reports. We therefore analyzed the incidence of peroneal nerve palsy, knee stability, and local recurrence following surgical treatment of benign proximal fibula tumors. We retrospectively reviewed the charts of 120 patients (121 tumors) with histologically confirmed aggressive benign tumors of the proximal fibula. There were 56 males and 64 females with an average age of 24 years (range, 2-64 years). The most common diagnosis was osteochondroma (38%) followed by giant cell tumor (19%). Pain (94%), palpable mass (39%), and peroneal nerve symptoms (12%) were the most common presenting symptoms. Of the 121 tumors, 56 (46%) underwent en bloc resection. The minimum followup was 2 years (mean, 9 years; range 2 to 49 years; median, 7.4 years). Postoperative complications included nine peroneal nerve palsies (six transient, three permanent), one deep venous thrombosis, and one wound dehiscence. No long-term knee instability was seen with repair of the lateral collateral ligament. Ten patients had recurrences, with 70% of local recurrences occurring in patients who underwent intralesional excision. Given the higher recurrence rate with curettage, patients with aggressive proximal fibula tumors benefit from en bloc resection. The overall morbidity is low, but postoperative permanent peroneal palsy remains a concern (3%).


Assessing quantum dimensionality from observable dynamics.
Using tools from classical signal processing, we show how to determine the dimensionality of a quantum system as well as the effective size of the environment's memory from observable dynamics in a model-independent way. We discuss the dependence on the number of conserved quantities, the relation to ergodicity and prove a converse showing that a Hilbert space of dimension D+2 is sufficient to describe every bounded sequence of measurements originating from any D-dimensional linear equations of motion. This is in sharp contrast to classical stochastic processes which are subject to more severe restrictions: a simple spectral analysis shows that the gap between the required dimensionality of a quantum and a classical description of an observed evolution can be arbitrary large.


Partial volume correction of brain perfusion estimates using the inherent signal data of time-resolved arterial spin labeling.
Quantitative perfusion MRI based on arterial spin labeling (ASL) is hampered by partial volume effects (PVEs), arising due to voxel signal cross-contamination between different compartments. To address this issue, several partial volume correction (PVC) methods have been presented. Most previous methods rely on segmentation of a high-resolution T1 -weighted morphological image volume that is coregistered to the low-resolution ASL data, making the result sensitive to errors in the segmentation and coregistration. In this work, we present a methodology for partial volume estimation and correction, using only low-resolution ASL data acquired with the QUASAR sequence. The methodology consists of a T1 -based segmentation method, with no spatial priors, and a modified PVC method based on linear regression. The presented approach thus avoids prior assumptions about the spatial distribution of brain compartments, while also avoiding coregistration between different image volumes. Simulations based on a digital phantom as well as in vivo measurements in 10 volunteers were used to assess the performance of the proposed segmentation approach. The simulation results indicated that QUASAR data can be used for robust partial volume estimation, and this was confirmed by the in vivo experiments. The proposed PVC method yielded probable perfusion maps, comparable to a reference method based on segmentation of a high-resolution morphological scan. Corrected gray matter (GM) perfusion was 47% higher than uncorrected values, suggesting a significant amount of PVEs in the data. Whereas the reference method failed to completely eliminate the dependence of perfusion estimates on the volume fraction, the novel approach produced GM perfusion values independent of GM volume fraction. The intra-subject coefficient of variation of corrected perfusion values was lowest for the proposed PVC method. As shown in this work, low-resolution partial volume estimation in connection with ASL perfusion estimation is feasible, and provides a promising tool for decoupling perfusion and tissue volume.